• Clinical and Experimental Reproductive Medicine
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• v.47 no.1
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• pp.68-76
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• 2020

Objective: Since sperm abnormalities are known to be a major reason for recurrent pregnancy loss (RPL), any defects in DNA structure and chromatin condensation can place embryos at risk in the early stage of development and implantation. As antioxidants such as vitamin C may play a protective role against the destruction of protamine genes in sperm chromatin, this study was conducted to evaluate the effects of vitamin C on chromatin and the expression of protamine genes in the male partners of couples with RPL. Methods: Twenty male partners of couples with RPL were selected as the intervention group and received vitamin C supplementation (250 mg daily for 3 months). Healthy fertile men (n = 20) were included as controls. Sperm chromatin, DNA integrity, and the expression levels of protamine genes were evaluated before and after treatment. Results: Significant differences were found in sperm morphology, protamine deficiency, and apoptosis between the two groups and before and after vitamin C administration. A significant change was found in mRNA levels of PRM1, PRM2, and the PRM1/PRM2 ratio after treatment. Conclusion: Daily oral administration of vitamin C may improve human sperm parameters and DNA integrity by increasing protamine gene expression levels in the male partners of couples with RPL. The beneficial effects of vitamin C supplementation as an antioxidant for the male partners of couples with RPL could lead to improved pregnancy outcomes in these cases.

• Animal Bioscience
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• v.37 no.1
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• pp.50-60
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• 2024

Objective: Testicular fat deposition has been reported to affect animal reproduction. However, the underlying mechanism remains poorly understood. The present study explored whether sperm meiosis and testosterone synthesis contribute to mouse testicular fat deposition-induced reproductive performance. Methods: High fat diet (HFD)-induced obesity CD1 mice (DIO) were used as a testicular fat deposition model. The serum hormone test was performed by agent kit. The quality of sperm was assessed using a Sperm Class Analyzer. Testicular tissue morphology was analyzed by histochemical methods. The expression of spermatocyte marker molecules was monitored by an immuno-fluorescence microscope during meiosis. Analysis of the synthesis of testosterone was performed by real-time polymerase chain reaction and reagent kit. Results: It was found that there was a significant increase in body weight among DIO mice, however, the food intake showed no difference compared to control mice fed a normal diet (CTR). The number of offspring in DIO mice decreased, but there was no significant difference from the CTR group. The levels of follicle-stimulating hormone were lower in DIO mice and their luteinizing hormone levels were similar. The results showed a remarkable decrease in sperm density and motility among DIO mice. We also found that fat accumulation affected the meiosis process, mainly reflected in the cross-exchange of homologous chromosomes. In addition, overweight increased fat deposition in the testis and reduced the expression of testosterone synthesis-related enzymes, thereby affecting the synthesis and secretion of testosterone by testicular Leydig cells. Conclusion: Fat accumulation in the testes causes testicular cell dysfunction, which affects testosterone hormone synthesis and ultimately affects sperm formation.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.10
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• pp.1417-1425
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• 2014

The selection of morphologically normal spermatozoa is critical to obtain high breeding performances in boar breeding farms and artificial insemination (AI) centers. Parameters for the selection of semen mainly include total sperm motility, concentration, and morphology. However, these primary parameters are often not reliable for discriminating between normal and abnormal, non-fertilizable spermatozoa. The present study was designed to compare the motion characteristics, fertilization ability using in vitro fertilization (IVF), and acrosome formation of the semen from boars having low (boar number 2012) and normal (boar number 2004 and 2023) breeding performances. The ultimate goal was to identify additional simple and easy criteria for the selection of normal sperm. There was no significant difference between boar 2004 and boar 2023 sperm total motility in computer assisted sperm analysis. However, boar number 2012 semen presented a significantly reduced population of rapid moving spermatozoa and an increased population of slow moving spermatozoa compared to boar numbers 2004 and 2023. Analysis of detailed motion characteristics revealed that sperm from boar number 2012 had significantly reduced motility in progressiveness, average path velocity, straight-line velocity (VSL), curvilinear velocity (VCL), straightness, and linearity. The assessment of the fertilizing ability by IVF also showed that sperm from boar number 2012 showed a fertility rate of 3.4%, whereas sperm from boar number 2023 had a fertility rate of 75.45%. Interestingly, most of the sperm nuclei were found on the peripheral area of the oocytes, suggesting that the sperm from boar number 2012 lacked penetration ability into the oocyte zonapellucida. The acrosome formation analysis using Pisum sativum agglutinin staining demonstrated that the sperm from boar number 2012 had a defect in acrosome formation. Consequently, primary parameters for selecting semen before AI such as motility are not sufficient to select normal and fertilizable spermatozoa. In conclusion, the present study suggests that the acrosome staining and detailed motion characteristics such as progressiveness, VCL, and VSL should be included in determining semen quality together with primary parameters for successful AI and high breeding performance in the swine industry.

• Biomedical Science Letters
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• v.10 no.1
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• pp.31-34
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• 2004

White blood cells (WBCs) are present in most human ejaculates, but abnormally high concentration of seminal leukocytes may reflect an underlying pathological condition. The World Health Organization (WHO) has defined leukocytospermia as status of more than $10^6$ WBC/mL of semen. The purpose of this study was firstly, to compare the outcomes between conventional IVF and ICSI in leukocytospermia, and secondly, to investigate whether ICSI may be an alternation treatment for patients with leukocytospermia. Total 346 cycles of conventional IVF and ICSI candidates underwent IVF cycles at Eulji Hospital Infertility Clinic. Semen Parameters including concentration, motility, morphology of spermatozoa and concentration of leukocytes were assessed from the raw ejaculates. There was no difference in sperm concentration, motility and morphology. The rates of fertilization and good embryo development from ICSI were significantly higher than those from conventional IVF in leukocytospermia (60.4% ＆ 32.5%, respectively for ICSI group and 44.4% ＆ 28.5%, respectively for IVF group, P<0.00l). The pregnancy rate after ICSI was also higher than that from conventional IVF (34.0% vs 29.1 %, P<0.05). These results indicate that the presence of seminal leukocyes (> 1$\times10^6$ WBC/mL of semen) is adversely related with fertilization, embryo development and pregnancy rate. Therfore the measurement of seminal leukocytes in routine semen analysis appears to be of prognostic value with regard to male fertilizing potential. In conclusion, it is suggested that ICSI is an alternative choice of treatment for patients with leukocytospermia.

• Korean Journal of Ecology and Environment
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• v.35 no.3 s.99
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• pp.232-236
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• 2002

In an effort to uncover the reproduction of Korean brook perch Coreoperca herzi testis anatomy and sperm morphology were studied. Fish samples were collected in the Sooypcheon river from May to October 2001. White-colored testes have wedgeshaped external morphology, and developed symmetrically in the dorsal cavity of the trunk. Isogenetic germ cells developed in the cyst located in seminiferous lobule. Each lobule showed significant asynchrony in the spermatogenic stage of the cyst. Sperm was 43 ${\mu}$m in length. The round head was 2.2 ${\mu}$m long. The middle piece developed beneath the head was 0.5 ${\mu}$m long. Tail was 40 ${\mu}$m in length. Coomassie brilliant blue (CBB) gave rise the intense staining in the apex of sperm head and middle piece, suggesting the possible development of acrosome.

• Journal of Veterinary Clinics
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• v.24 no.4
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• pp.577-583
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• 2007

It is important to obtain semen with good quality for efficient fertilization and pregnancy. To obtain these semen, various methods have been developed but most of these methods are time consuming and require costly equipment. Therefore, the objective of this research is to investigate the usability of column filtration system as quick and simple method to get sperm with better quality. Ejaculates were obtained from 5 dogs and analyzed with basic quality parameters before each filtration. Sperm concentration was adjusted to $5{\times}10^7/ml$ after dilution. The experimental groups were divided into non-filtered group(control) and filtered groups(glass wool, Sephadex 5% and Sephadex 20%). Ejaculates were filtered through each filter system and assessed by recovery rate of sperm, motility, normal morphology, CFDA/PI stain and plasma membrane integrity(hypo-osmotic swelling test, HOST). The lowest recovery rate of spermatozoa was recorded in glass wool filtration group, followed by 20% Sephadex filtration group(p<0.05). There was no significant difference between control(non-filtered) and 5% Sephadex filtration poop. Also, there was no significant difference of sperm motility assessed under light microscope among experimental groups. Morphological normality of canine spermatozoa was the highest in the glass wool filtration group and the lowest in the 5% Sephadex filtration group with no significant differences versus 20% Sephadex filtration and control group, respectively(p<0.05). Viability of canine sperm assessed by CFCA/PI staining was the highest in the glass wool filtration poop with no significant difference versus the control group, and the lowest in the 20% Sephadex filtration group with no significant difference versus 5% Sephadex filtration group, respectively(p<0.05). HOS values of canine sperm was the highest in the 20% Sephadex filtration group with no significant difference versus 5% Sephadex filtration group, and the lowest in the control poop with no significant difference versus glass wool filtration group, respectively(p<0.05). Therefore, these results indicated that filtration treatment for extended canine sperm would be useful method to get sperm with better quality by trapping the damaged sperm, consequently filter would be physical barrier against injured or immotile sperm.

• Clinical and Experimental Reproductive Medicine
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• v.43 no.4
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• pp.199-206
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• 2016

Objective: This study was carried out to investigate the correlations of the sperm DNA fragmentation index (DFI) with semen parameters and apoptosis, and to investigate the effects of density-gradient centrifugation (DGC) and magnetic-activated cell sorting (MACS) on reducing the proportion of sperm with DNA fragmentation and protamine deficiency. Methods: Semen analysis and a sperm DNA fragmentation assay were performed to assess the correlations between semen parameters and the DFI in 458 semen samples. Sperm with progressive motility or non-apoptosis were isolated by DGC or MACS, respectively, in 29 normozoospermic semen samples. The effects of DGC or MACS alone and of DGC and MACS combined on reducing the amount of sperm in the sample with DNA fragmentation and protamine deficiency were investigated. Results: The sperm DFI showed a significant correlation (r=-0.347, p< 0.001) with sperm motility and morphology (r=-0.114, p< 0.05) but not with other semen parameters. The DFI ($11.5%{\pm}2.0%$) of semen samples was significantly reduced by DGC ($8.1%{\pm}4.1%$) or MACS alone ($7.4%{\pm}3.9%$) (p< 0.05). The DFI was significantly further reduced by a combination of DGC and MACS ($4.1%{\pm}1.3%$, p< 0.05). Moreover, the combination of DGC and MACS ($1.6%{\pm}1.1%$, p< 0.05) significantly reduced the protamine deficiency rate of semen samples compared to DGC ($4.4%{\pm}3.2%$) or MACS alone ($3.4%{\pm}2.2%$). Conclusion: The combination of DGC and MACS may be an effective method to isolate high-quality sperm with progressive motility, non-apoptosis, high DNA integrity, and low protamine deficiency in clinical use.

• The Journal of Korean Medicine
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• v.25 no.4
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• pp.90-94
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• 2004

Objective : This study was conducted to investigate the effects of Panax Ginseng (인삼) on the sperm motility and spermatogenesis in the male rat. Methods : We used 8-week-old Sprague-Dawley rats, and administered the extract powder of Panax Ginseng to 5 rats (treated group) and normal saline (control group) once a day for 28 days. We isolated their testes surgically, then observed the change of the body weights before and after administration of Panax Ginseng extracts and normal saline. We observed the weight of the testes, epididymis, vascular gland, and prostate. Also, we examined the total, normal motile sperm concentration, and the concentration of testicular catalase and peroxidase. Results : We found that the concentration of normal, motile sperm in the testes of the Panax Ginseng group showed a significant difference compared with the control group. The angiogenesis of the seminiferous tubule was increased and the increasement of the number of spermatogonia, primary and secondary spermatocyte was observed in the Panax Ginseng group through a microscope. The body weight, the weight of the testes, epididymis, prostate and the concentration of testicular catalase and peroxidase were higher in the Panax Ginseng group but showed no significant difference. Conclusion : This study shows that Panax Ginseng may have an effect on the morphology and motility of sperm, the important factor in male fertility, and can promote the concentration of antioxidants, catalase and peroxidase, which is the important factor in spermatogenesis.

• The Journal of Korean Obstetrics and Gynecology
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• v.19 no.2
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• pp.49-61
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• 2006

Purpose : This study was undertaken to evaluate the effects of the different administration duration of Ginseng Radix Alba extract solution on the spermatogenic abilities. Methods : We used the 2-month-old mice and administered Ginseng Radix Alba extract solution 0.3mg/g/day for 30, 60, 90 and 120 days. The control group was administered the normal saline in the same way. We examined the number of total, motile and normal sperm from the cauda epididymis, the activities of sperm hyaluronidase. Also we observed changes of isolated testis before and after administration of Ginseng Radix Alba extract solutions in the mice. And we compared to the testicular tissue especially seminiferous tubules between control and treated group by histochemical methods. Results : The significant differences were observed in the concentration of total sperm and normality of spermatozoa of the Ginseng Radix Alba extract solution administered groups compared to the control group in 60, 90 and 120days groups. The significant differences were observed the motility of the Ginseng Radix Alba extract solution administered groups compared to the control group in 90 days group respectively. In the histological analysis of the testicular tissues, the enlargement of testicular lobe diameter and apparent vasculogenesis between testicular lobes were observed in the Ginseng Radix Alba extract solution administered groups compared to the control group, respectively. but the activity of hyaluronidase was not significantly increased in the Ginseng Radix Alba extract solution administered groups Conclusion : This study shows that Ginseng Radix Alba has the beneficial effect on the concentration, morphology and motility of sperm especially in 90 days administration group. We can suggest that Ginseng Radix Alba extract solution be useful for the treatment of male sexual dysfunctions and infertility.

• The Journal of Korean Obstetrics and Gynecology
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• v.18 no.4
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• pp.24-35
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• 2005

Purpose : This study was conducted to investigate the effects of Cynomorii Herba (鎖陽) on　the reproductive competence in male mice.　Methods ：　We used 8-week-old Spraque-Dawley rats and administered the extract powder of Cynomorii Herba to 5 rats and normal saline to control group once in a day for 28 days. Then we observed the changes of body weight before and after administration of Cynomorii Herba extracts and normal saline. We isolated their testis surgically and observed the weight of testis, epididymis, vascular gland and prostate. Also we examined the total, normal and motile sperm concentration, the concentration of testicular catalase, peroxidase and configuration of testicular tissue before and after administration of Cynomorii Herba extracts and normal saline. Results : We found that the concentration of total, normal, motile sperm in testis of Cynomorii Herba treated group shows significant difference compared with the control group. The body weight, the weight of testis, epididymis and the concentration of testicular catalase and peroxidase were higher in the Cynomorii Herba treated group, on the other hand the weight of vascular gland was lower in the Cynomorii Herba. But the contents of above statement showed no significant difference. The weight of　prostate showed lower in the control group statistically than that of the Cynomorii Herba treated group. We observed the seminiferous tubules taken shape minutely and the number of normal sperm increased in Cynomorii Herba treated group's testicular tissue. Conclusion This study shows that Cynomorii Herba may have an effect on the concentration, morphology and motility of sperm, the important factor in male fertility.